Zhang Ying, Xu Bo, Kinoshita Naohiko, Yoshida Yutaka, Tasaki Masayuki, Fujinaka Hidehiko, Magdeldin Sameh, Yaoita Eishin, Yamamoto Tadashi
Department of Structural Pathology, Institute of Nephrology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan; Biofluid Biomarker Center (BB-C), Institute for Research Collaboration and Promotion, Niigata University, Niigata, Japan.
Department of Structural Pathology, Institute of Nephrology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan.
J Proteomics. 2015 Jun 18;123:89-100. doi: 10.1016/j.jprot.2015.03.024. Epub 2015 Mar 30.
Since glomerular sclerosis frequently accompanies various glomerular diseases at the end stages, it is challenging to differentiate ubiquitous biological processes underlying this pathology from those critically involved in specific diseases. Furthermore, in-depth proteomic profile of human glomerular sclerosis remains limited. In this study, human glomeruli with intermediate (i-GS) and advanced (GS) sclerotic lesions, which were excluded from specific renal diseases and assumed to be aging-related, were laser captured from macroscopically normal cortex distant from urological carcinoma, and subjected to label-free quantitative proteomic analysis. We explicate an evident increase of membrane attack complex in i-GS and GS with an up-going tendency, which is accompanied by increasing of inhibitory regulators of alternative and terminal pathways. GO annotation and IPA pathway analysis agree to these results. Proteomic findings are validated by immunohistochemical studies which indicate that alternative and terminal pathways are positively involved in the glomerular sclerosis seen in distinct renal diseases. Furthermore, proteomic analysis also demonstrates remarkable increases of complement factor B in GS and TGF-ß1 in both GS and i-GS. Identification of complement factor B implicates that on-site activation of alternative pathway may occur in injured glomeruli and stepwise increase of TGF-ß1 suggests its contribution to the progression of glomerulosclerosis.
This study provides in-depth quantitative proteomic profiles of human glomeruli with intermediate and advanced sclerotic lesions. It reveals that the over-expression of alternative and terminal pathway components is significantly involved in human glomerulosclerosis seen in distinct renal diseases. Proteomic identification of the increased TGF-ß1 provides supporting evidence for the role of podocyte apoptosis leading to human glomerulosclerosis.
由于肾小球硬化在终末期常伴随各种肾小球疾病，因此很难将这种病理状态下普遍存在的生物学过程与特定疾病中关键涉及的过程区分开来。此外，人类肾小球硬化的深度蛋白质组学特征仍然有限。在本研究中，从远离泌尿系统癌的宏观正常皮质中激光捕获具有中度（i-GS）和重度（GS）硬化病变的人类肾小球，这些病变被排除在特定肾脏疾病之外，被认为与衰老相关，并进行无标记定量蛋白质组学分析。我们发现i-GS和GS中膜攻击复合物明显增加且呈上升趋势，同时替代途径和终末途径的抑制调节因子也增加。基因本体注释（GO注释）和IPA通路分析证实了这些结果。蛋白质组学研究结果通过免疫组织化学研究得到验证，免疫组织化学研究表明替代途径和终末途径积极参与不同肾脏疾病中的肾小球硬化。此外，蛋白质组学分析还显示GS中补体因子B显著增加，GS和i-GS中转化生长因子β1（TGF-β1）均显著增加。补体因子B的鉴定表明替代途径可能在受损肾小球中发生原位激活，TGF-β1的逐步增加表明其对肾小球硬化进展的作用。
本研究提供了具有中度和重度硬化病变的人类肾小球的深度定量蛋白质组学特征。它揭示了替代途径和终末途径成分的过表达显著参与不同肾脏疾病中的人类肾小球硬化。蛋白质组学鉴定出增加的TGF-β1为足细胞凋亡导致人类肾小球硬化的作用提供了支持证据。
